WO1994012520A1 - Segment de liaison pour polypeptides fusionnes lies - Google Patents

Segment de liaison pour polypeptides fusionnes lies Download PDF

Info

Publication number
WO1994012520A1
WO1994012520A1 PCT/US1993/011138 US9311138W WO9412520A1 WO 1994012520 A1 WO1994012520 A1 WO 1994012520A1 US 9311138 W US9311138 W US 9311138W WO 9412520 A1 WO9412520 A1 WO 9412520A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
fusion polypeptide
linked fusion
chain
peptide linker
Prior art date
Application number
PCT/US1993/011138
Other languages
English (en)
Inventor
Marc D. Whitlow
David Fipula
Original Assignee
Enzon, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Enzon, Inc. filed Critical Enzon, Inc.
Priority to AU56701/94A priority Critical patent/AU5670194A/en
Publication of WO1994012520A1 publication Critical patent/WO1994012520A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3046—Stomach, Intestines
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S930/00—Peptide or protein sequence
    • Y10S930/01—Peptide or protein sequence
    • Y10S930/31—Linker sequence

Definitions

  • the present invention relates to linked fusion polypeptides derived from single and multiple chain proteins.
  • the invention relates to the linker peptide essential for bridging the polypeptide constituents that comprise the linked fusion polypeptide.
  • the gene sequence coding for a desired natural protein is isolated, replicated (cloned) and introduced into a foreign host such as a bacterium, a yeast (or other fungi) or a mammalian cell line in culture, with appropriate regulatory control signals. When the signals are activated, the gene is transcribed and translated, and expresses the desired protein. In this manner, such useful biologically active materials as hormones, enzymes and antibodies have been cloned and expressed in foreign hosts.
  • a foreign host such as a bacterium, a yeast (or other fungi) or a mammalian cell line in culture, with appropriate regulatory control signals.
  • the signals When the signals are activated, the gene is transcribed and translated, and expresses the desired protein.
  • useful biologically active materials as hormones, enzymes and antibodies have been cloned and expressed in foreign hosts.
  • One of the problems with this approach is that it is limited by the "one gene, one polypeptide chain" principle of molecular biology.
  • a genetic sequence codes for a single polypeptide chain.
  • Many biologically active polypeptides are aggregates of two or more chains.
  • antibodies are three-dimensional aggregates of two heavy and two light chains.
  • large enzymes such as aspartate transcarbamylase, for example, are aggregates of six catalytic and six regulatory chains, these chains being different.
  • Aspartate transcarbamylase for example, are aggregates of six catalytic and six regulatory chains, these chains being different.
  • genes can be expressed in separate hosts. The resulting polypeptide chains from each host would then have to be reaggregated and allowed to refold together in solution.
  • the two or more genes coding for the two or more polypeptide chains of the aggregate could be expressed in the same host simultaneously, so that refolding and reassociation into the native structure with biological activity will occur after expression.
  • This approach necessitates expression of multiple genes in a single host. Both of these approaches have proven to be inefficient. Even if the two or more genes are expressed in the same organism it is quite difficult to get them all expressed in the required amounts.
  • Antibodies are immunoglobulins typically composed of four polypeptides; two heavy chains and two light chains. Genes for heavy and light chains have been introduced into appropriate hosts and expressed, followed by reaggregation of these individual chains into functional antibody molecules (see, for example, Munro, Nature 312:591 (1984); Morrison, S.L., Science 229:1202' (1985); and Oi et al, BioTechniques 4:214 (1986); Wood et al., Nature 374:446-449 (1985)). Antibody molecules have two generally recognized regions in each of the heavy and light chains.
  • variable region which is responsible for binding to the specific antigen in question
  • constant region which is responsible for biological effector responses such as complement binding, etc.
  • the constant regions are not necessary for antigen binding.
  • the constant regions have been separated from the antibody molecule, and biologically active (i.e., binding) variable regions have been obtained.
  • variable regions of a light chain (VJ and a heavy chain (V H ) together form the structure responsible for an antibody's binding capability.
  • Antibodies may be cleaved to form fragments, some of which retain their binding ability.
  • One such fragment is the "Fv" fragment, which is composed of the terminal binding portions of the antibodies.
  • the Fv comprises two complementary subunits, the V L and V H , which in the native antibody compose the binding domains.
  • the Fv fragment of an antibody is probably the minimal structural component which retains the binding characteristics of the parent antibody.
  • the limited stability at low protein concentrations of the Fv fragments may be overcome by using an artificial peptide linker to join the variable domains of an Fv.
  • the resulting single-chain Fv (hereinafter "sFv") polypeptides have been shown to have binding affinities equivalent to the monoclonal antibodies
  • MAbs from which they were derived (Bird et al., Science 242:423 (1988)).
  • catalytic MAbs may be converted to a sFv form with retention of catalytic characteristics (Gibbs et al., Proc. Na l. Acad. Sci., USA 88:4001 (1991)).
  • sFv single-chain Fv
  • Single-chain Fv polypeptides are small proteins with a molecular weight around 27 kd, which lack the constant regions of 50 kd Fab fragments or 150 kd immunoglobulin antibodies bearing gamma chains (IgG). Like a Fab fragment, and unlike an IgG, an sFv polypeptide contains a single binding site.
  • sFv polypeptides Due to their small size, sFv polypeptides clear more rapidly from the blood and penetrate more rapidly into tissues (Colcher, et al., J. Natl. Cancer Inst. 82:1191 (1990); Yokota et al., Cancer Research 52:3402 (1992)). Due to lack of constant regions, sFv polypeptides are not retained in tissues such as the liver and kidneys. Due to the rapid clearance and lack of constant regions, sFv polypeptides will have low immunogenicity. Thus, sFv polypeptides have applications in cancer diagnosis and therapy, where rapid tissue penetration and clearance are advantageous.
  • sFv polypeptides can be engineered with the two variable regions derived from a MAb.
  • the sFv is formed by ligating the component variable domain genes with an oligonucleotide that encodes an appropriately designed linker polypeptide.
  • the linker bridges the C-terminus of the first V region and the N-terminus of the second V region.
  • sFv polypeptides offer a clear advantage over MAbs because they do not have the constant regions derived from their biological source, which may cause antigenic reaction against the MAb.
  • Single-chain immunotoxins have been produced by fusing a cell binding sFv with Pseudomonas exotoxin (Chaudhary et al., Nature 339:394 (1989)). Recently, a single-chain immunotoxin was shown to cause tumor regression in mice (Brinkmann et al., Proc. Natl. Acad. Sci. USA 88:8616 (1991)).
  • TRY40 is a double linker with 3- and 7-amino acid sequences comprising the linkers.
  • the sequences are PGS and IAKAFKN (see page 8, Table 1 for a description of the single letter amino acid code used herein).
  • TRY59 is an 18-residue single linker having the sequence KESGSVSSEQLAQFRSLD (SEQ. ID No. 2).
  • TRY 61 is a 14-residue single linker having the sequence VRGSPAINVAVHVF (SEQ. ID No. 3).
  • TRY104b is a 22-residue single linker constructed primarily of a helical segment from human hemoglobin. The sequence is AQGTLSPADKTNV KAAWGKVMT (SEQ. ID No. 4).
  • GGSGGSGGVD SEQ. ID No. 5
  • the final bispecific single-chain polypeptide is called Janusin, and targets cytotoxic lymphocytes on HIV- infected cells.
  • Linkers previously used for sFvs and other polypeptides suffer from proteolytic attack, rendering them less stable and prone to dissociation. They also suffer from inordinate aggregation at high concentrations, making them susceptible to concentration in the liver and kidneys. Therefore, .there is a need for more stable linkers that are resistant to proteolytic attack and less prone to aggregation.
  • the invention is directed to a linked fusion polypeptide comprising polypeptide constituents connected by a novel peptide linker.
  • the novel peptide linker comprises a sequence of amino acids numbering from about 2 to about 50 having a first end connected to a first protein domain, and having a second end connected to a second protein domain, wherein the peptide comprises at least one proline residue within the sequence, the proline being positioned next to a charged amino acid, and the charged amino acid-proline pair is positioned within the peptide linker to inhibit proteolysis of said polypeptide.
  • the invention is also directed to a novel peptide linker comprising the amino acid sequence:
  • the invention also relates to sFvs wherein the linker linking V H and V L regions is the peptide linker as herein described, preferably comprising from about 10 to about 30 amino acids, and more preferably comprising at least 18 amino acids.
  • the invention also relates to genetic sequences encoding linked fusion polypeptides containing the novel peptide linker herein described, methods of making such linked fusion polypeptides, and methods of producing such linked fusion polypeptides via recombinant DNA technology.
  • Figure 1 is a set of two graphs depicting the proteolytic susceptibility of the CC49/212 and CC49/218 sFv proteins when exposed to subtilisin BPN' (Panel A) or trypsin (Panel B). The fraction of sFv remaining intact was determined by reverse phase HPLC. The CC49/212 sFv is shown in open circles and the CC49/218 is shown in closed squares. There was no measurable degradation of the CC49/218 sFv.
  • Figure 2 is a graph depicting the results of a competition radioimmunoassay (RIA) in which unlabeled CC49/212 single-chain Fv (open squares), CC49/218 single-chain Fv (closed diamonds) or MOPC-21 IgG (+) competed against a CC49 IgG radiolabeled with ,25 I for binding to the TAG-72 antigen on a human breast carcinoma extract.
  • RIA radioimmunoassay
  • Figure 3 is the amino acid (SEQ. ID No. 12) and nucleotide (SEQ. ID No. 11) sequence of the linked fusion polypeptide comprising the 4-4-20 V L region connected through the 217 linker to the CC49 V H region.
  • Figure 4 is the amino acid (SEQ. ID No. 14) and nucleotide (SEQ. ID No. 13) sequence of the linked fusion polypeptide comprising the CC49 V L region connected through the 217 linker polypeptide to the 4-4-20 V H region.
  • Figure 5 is a chromatogram depicting the purification of CC49/4-4-20 heterodimer Fv on a cation exchange high performance liquid chromatographic column. The column is a PolyCAT A aspartic acid column (Poly LC, Columbia, MD). The heterodimer Fv is shown as peak 5, eluting at 30.10 min.
  • Figure 6 is a coomassie-blue stained 4-20% SDS-PAGE gel showing the proteins separated in Figure 5. Lane 1 contains the molecular weight standards. Lane 3 contains the starting material before separation. Lanes 4-8 contain fractions 2, 3, 5, 6 and 7, respectively. Lane 9 contains purified CC49/212.
  • Figure 7 is a chromatogram used to determine the molecular size of fraction 2 from Figure 5.
  • a TSK G3000SW gel filtration HPLC column was used (Toyo Soda, Tokyo, Japan).
  • Figure 8 is a chromatogram used to determine the molecular size of fraction 5 from Figure 5.
  • a TSK G3000SW gel filtration HPLC column was used (Toyo Soda, Tokyo, Japan).
  • Figure 9 is a chromatogram used to determine the molecular size of fraction 6 from Figure 5.
  • a TSK G3000SW gel filtration HPLC column was used (Toyo Soda, Tokyo, Japan).
  • Figure 10 shows a Scatchard analysis of the fluorescein binding affinity of die CC49/4-4-20 heterodimer Fv (fraction 5 in Figure 5).
  • Figure 11 is a graphical representation of three competition enzyme- linked immunosorbent assays (ELISA) in which unlabeled CC49/4-4-20 Fv (closed squares) CC49/212 single-chain Fv (open squares) and MOPC-21 IgG ( • +) competed against a biotin-labeled CC49 IgG for binding to the TAG-72 antigen on a human breast carcinoma extract.
  • ELISA enzyme- linked immunosorbent assays
  • a protein is a biological molecule which consists primarily of one or more polypeptides.
  • a protein consisting of a single polypeptide is referred to herein as a single chain protein.
  • a protein consisting of more than one polypeptide is referred to herein as a multi-chain protein, with the term chain being synonymous with the term polypeptide.
  • Polypeptide As referred to herein, a polypeptide is a linear, single chain polymer of multiple amino acids linked through their amino and carboxylate groups by peptide bonds.
  • a polypeptide may form a single chain protein by itself or, in association with other polypeptides, form a multi-chain protein.
  • a polypeptide may also be a fragment of a single chain protein or a fragment of one of the chains of a multi-chain protein.
  • a linked fusion polypeptide is a polypeptide made up of two smaller polypeptide constituents, each constituent being derived from a single chain protein or a single chain of a multi-chain protein, where the constituents are combined in a non-naturally occurring arrangement using a peptide linker.
  • Linked fusion polypeptides mimic some or all of the functional aspects or biological activities of the protein(s) from which their polypeptide constituents are derived.
  • the constituent at the amino terminal portion of the linked fusion polypeptide is referred to herein as the first polypeptide.
  • the constituent at the carboxy terminal portion of the linked fusion polypeptide is referred to herein as the second polypeptide.
  • non-naturally occurring arrangement is meant an arrangement which occurs only through in vitro manipulation of either the polypeptide constituents themselves or the nucleic acids which encode them.
  • a peptide linker or linker is a polypeptide typically ranging from about 2 to about 50 amino acids in length, which is designed to facilitate the functional connection of two polypeptides into a linked fusion polypeptide.
  • the term functional connection denotes a connection that facilitates proper folding of the polypeptides into a three dimensional structure that allows the linked fusion polypeptide to mimic some or all of the functional aspects or biological activities of the protein(s) from which its polypeptide constituents are derived.
  • connection In cases such as sFv polypeptides where the linker is used to make a single chain derivative of a multi-chain protein, to achieve the desired biological activity the appropriate three dimensional structure will be one that mimics the structural relationship of the two polypeptide constituents in the native multi-chain protein.
  • the term functional connection also denotes a connection that confers a degree of stability required for the resulting linked fusion polypeptide to function as desired.
  • a charged amino acid is a biologically derived amino acid which contains a charge at neutral pH.
  • Charged amino acids include the negatively charged amino acids Aspartic acid (D) and Glutamic acid (E) as well as positively charged amino acids Histidine (H), Lysine (K), and Arginine (R).
  • Immunoglobulin superfamily As referred to herein, the immunoglobulin superfamily is the family of proteins containing one or more regions that resemble the variable or constant regions of an immunoglobulin, or fundamental structural units (i.e., domains) found within these regions.
  • the resemblance referred to is in terms of size, amino acid sequence, and presumably three dimensional structure.
  • immunoglobulin superfamily typically mediate non-enzymatic intercellular surface recognition and include, but are not limited to, CD1, CD2, CD3, CD7, CD8, CD28 class I and II histocompatibility molecules, Beta-2 microglobulin, lymphocyte function associated antigen-3 (LFA-3), FC ⁇ receptor, Thy-1, T cell receptor, polyimmunoglobulin receptor, neuronal cell adhesion molecule, myelin associated glycoprotein, P 0 myelin, carcinoembryonic antigen, platelet derived growth factor receptor, colony stimulating factor-1 receptor, link protein of basement membrane, and o_,
  • T cell Receptor As referred to herein, T cell receptor is a member of the immunoglobulin superfamily that resides on the surface of T lymphocytes and specifically recognizes molecules of the major histocompatibility complex, either alone or in association with foreign antigens.
  • Immunoglobulin As referred to herein, an immunoglobulin is a multi-chain protein with antibody activity typically composed of two types of polypeptides, referred to as heavy and light chains.
  • the heavy chain is larger than the light chain and typically consists of a single variable region, three or four constant regions, a carboxy-terminal segment and, in some cases, a hinge region.
  • the light chain typically consists of a single variable region and a single constant region.
  • an antibody is an immunoglobulin that is produced in response to stimulation by an antigen and that reacts specifically with that antigen.
  • Antibodies are typically composed of two identical heavy and two identical light polypeptide chains, held together by interchain disulfide bonds.
  • Single chain Fv polypeptide As referred to herein, a single chain Fv polypeptide (sFv) is a linked fusion polypeptide composed of two variable regions derived from the same antibody, connected by a peptide linker. An sFv is capable of binding antigen similar to the antibody from which its variable regions are derived. An sFv composed of variable regions from two different antibodies is referred to herein as a mixed sFv.
  • variable domain (V, ) to extend from residue 1 to residue 107 for the lambda light chain, and to residue 108 for kappa light chains, and the variable domain of the heavy chain (V H ) to extend from residue 1 to residue 113.
  • V L is the N-terminal domain followed by the linker and V H (a V L -
  • V H is the N-terminal domain followed by the linker and V L (V H -Linker-V L construction). Alternatively, multiple linkers have also been used.
  • V H -Linker-V L construction Several types of sFv proteins have been successfully constructed and purified, and have shown binding affinities and specificities similar to the antibodies from which they were derived.
  • the Fv domains have been selected from the group of monoclonal antibodies known by their abbreviations in the literature as 26-10, MOPC 315, 741F8, 520C9, McPC 603, D1.3, murine phOx, human phOx, RFL3.8 sTCR, 1A6, Sel55-4, 18-2-3, 4-4-20, 7A4-1, B6.2, CC 49, 3C2, 2c, MA-15C5/K, 2 G 0 , Ox, etc. (see references previously cited as disclosing Fv analog polypeptides).
  • the Fv's are derived from the variable regions of the corresponding monoclonal antibodies (MAbs).
  • Linkers have also been used to join non-antibody polypeptides, as evidenced by Soo Hoo et al, Proc. Natl Acad. Sci. USA 89:4759-4763
  • linkers according to the invention will be useful for connecting polypeptides derived from any protein.
  • the order in which the polypeptides are connected i.e., which is nearer the amino or carboxy terminus of the linked fusion polypeptide should, where possible, reflect the relationship of the polypeptides in their native state.
  • the polypeptide derived from the first chain should be positioned near the amino-terminal portion of the linked fusion polypeptide and the polypeptide derived from the second chain should be positioned near the carboxy-terminal portion.
  • linkers according to the invention will be applicable to any multi-chain protein or protein complex including, but not limited to, members of the immunoglobulin superfamily, enzymes, enzyme complexes, ligands, regulatory proteins, DNA-binding proteins, receptors, hormones, etc.
  • proteins or protein complexes include, but are not limited to, T cell receptors, insulin, RNA polymerase, Myc, Jun, Fos, glucocorticoid receptor, thyroid hormone receptor, acetylcholine receptor, fatty acid synthetase complex, hemoglobin, tubulin, myosin, 3-Lactoglobulin, aspartate transcarbamoylase, malic dehydrogenase, glutamine synthetase, hexokinase, glyceraldehyde-phosphate dehydrogenase, glycogen phosphorylase, tryptophan synthetase, etc.
  • non-polypeptide biochemical moieties including, but not limited to, toxins, drugs, radioisotopes, etc. may be added to, or associated with, the linked fusion polypeptides to achieve a desired effect, such as labeling or conferring toxicity.
  • the preferred length of the peptide linker should be from 2 to about 50 amino acids. In each particular case, the preferred length will depend upon the nature of the polypeptides to be linked and the desired activity of the linked fusion polypeptide resulting from the linkage. Generally, the linker should be long enough to allow the resulting linked fusion polypeptide to properly fold into a conformation providing the desired biological activity. Where conformational information is available, as is the case with sFv polypeptides discussed below, the appropriate linker length may be estimated by consideration of the 3-dimensional conformation of the substituent polypeptides and the desired conformation of the resulting linked fusion polypeptide.
  • linker length may be empirically determined by testing a series of linked fusion polypeptides with linkers of varying lengths for the desired biological activity.
  • Linkers of the invention used to construct sFv polypeptides are designed to span the C terminus of V L (or neighboring site thereof) and the N terminus of V H (or neighboring site thereof) or between the C terminus of V H and d e N terminus of V L .
  • the linkers used to construct sFv polypeptides have between 10 and 30 amino acid residues.
  • the linkers are designed to be flexible, and it is recommended that an underlying sequence of alternating Gly and Ser residues be used. To enhance the solubility of the linker and its associated single chain
  • three charged residues may be included, two positively charged lysine residues (K) and one negatively charged glutamic acid residue (E).
  • K positively charged lysine residues
  • E negatively charged glutamic acid residue
  • one of the lysine residues is placed close to the N-terminus of V H , to replace the positive charge lost when forming the peptide bond of the linker and the V H .
  • linker lengths of equal to or greater than 18 residues reduce aggregation. This becomes important at high concentrations, when aggregation tends to become evident. Thus, linkers having 18 to 30 residues are preferred for sFv polypeptides.
  • Another property that is important in engineering an sFv polypeptide, or any other linked fusion polypeptide, is proteolytic stability.
  • the 212 linker (Pantoliano et al, Biochemistry 30: 10117 (1991)) is susceptible to proteolysis by subtilisin BPN'. The proteolytic clip in the 212 linker occurs between Lys8 and Ser9 of the linker (see Table 2).
  • proline residue in the peptide linker of the present invention inhibits the charge- transfer intermediate that is essential to the hydrolysis of the amide bond joining the two amino acid residues clipped apart by serine proteases.
  • Table 2 shows two of the claimed linkers (217 and 218) and two of the prior art linkers (202 ' and 212) for illustration.
  • the 217 linker contains a lysine-proline pair at positions 6 and 7, thus rendering the linker less susceptible to proteolysis.
  • the 218 linker demonstrates less aggregation, proteolytic stability, and the necessary flexibility and solubility to result in a functional linker for sFv proteins.
  • the anti-fluorescein sFvs 4-4-20/202', 4-4-20/212 and 4-4-20/216 had affinities of 0.5 x 10 9 M "1 , 1.0 x 10 9 M 1 , and 1.3 x 10° M” 1 , respectively using the fluorescence quenching assay.
  • 4-4-20 sFvs was that the prior art 212 linker was proteolytically susceptible. It was possible to produce crystals of the 4-4-20/212 sFv only after it had been treated with subtilisin BPN', a serine protease. When 4-4-20/212 sFv and subtilisin BPN' were mixed in a 5000 to 1 ratio, the 27 kD band of the sFv was converted into two bands that ran just below the 14 kD marker on the
  • Placement of the proline next to a charged residue is also preferred.
  • the sequence of proline and a charged amino acid residue should be maintained with the charged residue before (i.e. , on the amino-terminus side of) the proline.
  • a lysine-proline pair is located at the cleavage site, replacing die susceptible amide bond that was hydrolyzed.
  • arginine may be used as the charged residue.
  • a second guiding consideration in designing the linker of the invention is that a linker with reduced aggregation is preferable. As described above, the 18-residue 216 linker shows reduced aggregation as compared to the 14- residue 212 linker.
  • the first eleven residues of the 216 linker are identical to the 212 linker, including the proteolytically-susceptible peptide bond between Lys 8 and Ser 9. Thus, it is believed that the extra four residues contribute to the lowered aggregation. Linkers with 18 or more residues are thus preferred.
  • Positioning the proline at the proper place in the linker sequence to inhibit proteolysis is accomplished by determining the points of proteolytic attack in the susceptible sequence.
  • a protease such as subtilisin BPN' is contacted with the candidate linker. Cleavage can then be determined by sequencing the resulting peptides, which will also reveal the cleavage point or points, if any. Any protease may be used, and selection will be guided by consideration of the environment the linker is to encounter in actual use.
  • DNA molecules such as isolated genetic sequences or plasmids or vectors encoding linked fusion polypeptides with the peptide linker of the invention.
  • the DNA sequence for the linked fusion polypeptide can be chosen so as to optimize production in organisms such as bacteria or veast. Recombinant hosts as well as methods of using them to produce single chain proteins by expression, are also provided herein.
  • Patent No. 4,946,778, which is fully incorporated herein by reference.
  • the rates of aggregation of the sFv polypeptides were determined at room temperature in 60 mM MOPS, pH 7.0 at various concentrations using Gel Filtration HPLC Chromatography. 10 to 50 ⁇ l samples were injected onto a Waters HPLC system with 7.8 mm x 300 mm TSK G3000SW column (Toso Haas, Tokyo, Japan). The column had been previously equilibrated and the samples were eluted using 50 mM MOPS, 100 mM NaCl, buffer pH 7.5 at a flow rate of 0.5 ml/min. The data was collected on a Macintosh SE (Apple Computer, Cupertino, CA) running the
  • MAb CC49 and CC49 sFv polypeptides were labeled with Na I 5 I using Iodo-Gen (Pierce Chemical Co., Rockford, IL) as previously reported (Milenic, D., et al, Cancer Res. 57:6363-6371 (1991)).
  • the CC49 sFv polypeptides were labeled with the lutetium complex of the macrocyclic bifunctional coordinator PA-DOTA (Cheng et al , European Patent Application No. 353,450). 20 ⁇ l of a 1 mM solution of SCN-PA- DOTA in water was mixed with equal volumes of the I77 Lu(NO 3 ) 3 solution and 1 M HEPES buffer pH 7.0 and left at room temperature for five minutes.
  • 177 Lu in 0.05 N HC1 was obtained from the University of Missouri Research Reactor (Columbia, MO). The reaction mixture was processed over a PRP-1 reverse-phase cartridge (Hamilton Co., Reno, NV) which had been equilibrated with 10% acetonitrile in 20 mM sodium carbonate, pH 9.5. 177 Lu-SCN-PA-DOTA was eluted with acetonitrile/carbonate buffer (1:2) and a 60 ⁇ l fraction containing the radioactive chelate was used.
  • the half-life of the CC49/212 sFv treated with subtilisin or trypsin is 122.8 min and 195.7 min, respectively (see Figure 1).
  • the 218 linker had significantly improved protease resistance, for in the 48 hour period digestion of the CC49/218 sFv was not detectable using either subtilisin or trypsin.
  • a competition radioimmunoassay (RIA) was set up in which a CC49 IgG labeled with 125 I was competed against the unlabeled CC49 sFvs for binding to TAG-
  • the binding affinities for the TAG-72 antigen of the CC49/212 and CC49/218 sFv polypeptides were checked.
  • the CC49/218 sFv showed about a 4-fold lower affinity than the CC49/212 sFv (see Figure 2).
  • the lower affinity of the CC49/218 sFv could be in part due to the higher degree of aggregation of the CC49/212 sFv sample.
  • We have shown previously that the dimeric forms of CC49 (IgG and F(ab0 2 ) compete with a ten-fold higher affinity than do die monovalent forms (Fab and sFv) (Milenic, D., et al, Cancer Res. 57:6363-6371 (1991)). Since aggregates are multivalent it seems likely that they would have high affinity.
  • the rates of aggregation of the CC49/212 and CC49/218 sFv polypeptides were determined at room temperature (22°C) at various concentrations.
  • the CC49/212 sFv showed 80-fold faster accumulation of aggregates than did the CC49/218 sFv, at concentrations around 1.5 mg/ml (see Table 3). At 0.5 mg/ml this difference increased to 1600-fold.
  • the aggregation of both proteins showed a concentration dependence. The higher the concentration the higher the levels of aggregation that were seen.
  • mice Female athymic nude mice (nu/nu), obtained from Charles River
  • the biodistribution of the ,77 Lu labeled CC49/212 and CC49/218 sFv polypeptides was determined at various times in athymic nude mice bearing the two-week old human colon carcinomas. Of the six tissues examined, three tissues showed significant differences between the CC49/212 and CC49/218 sFvs (see Table 4). The spleen and the liver showed three- to four-fold higher accumulations of the CC49/212 sFv compared to the CC49/218 sFv. At the 24 and 48 hour time points the CC49/212 sFv showed a 60% higher accumulation at the tumor. The other three tissues (blood, kidney and lung) show little or no differences.
  • the higher level of CC49/212 sFv accumulation in the spleen and liver is likely due to the higher degree of aggregation of the sample injected. Both the spleen and liver metabolize the sFv polypeptides, but due to the higher degree of aggregation of the CC49/212 sFv higher uptake and accumulation of the 177 Lu radiolabel in these tissues is seen.
  • the higher levels of CC49/212 sFv in the tumor at later times may be due to the increased avidity of the aggregates.
  • the very high levels of accumulation of both sFv polypeptides in the kidneys probably reflects the catabolism of the protein in the kidneys, with subsequent retention of the l77 Lu (Schott et al, submitted).
  • the goals of this experiment were to produce, purify and analyze for activity a new heterodimer Fv that would bind to botii fluorescein and the pan- carcinoma antigen TAG-72.
  • the design consisted of two polypeptide chains, which associated to form the active heterodimer Fv. Each polypeptide chain can be described as a mixed single-chain Fv (mixed sFv).
  • the first mixed sFv (GX 8952) comprised a 4-4-20 variable light chain (VJ and a CC49 variable heavy chain (V H ) connected by a 217 polypeptide linker (Figure 3).
  • the second mixed sFv (GX 8953) comprised a CC49 V L and a 4-4-20 V H connected by a 217 polypeptide linker ( Figure 4).
  • the sequence of the 217 polypeptide linker is shown in Table 2.
  • the supernatant was discarded after centrifugation and die pellets resuspended in 2.5 liters of lysis/wash buffer at 4°C. This suspension was centrifuged for 45 minutes at 8000 rpm with the Dupont GS-3 rotor. The supernatant was again discarded and the pellet weighed. The pellet weight was 136.1 gm.
  • the anti-fluorescein activity was checked by a 40% quenching assay, and the amount of active protein calculated. 150mg total active heterodimer Fv was found by the 40% quench assay, assuming a 54,000 molecular weight.
  • the filtered sample of heterodimer was dialyzed, using a Pellicon system containing 10,000 dalton MWCO membranes, with dialysis buffer
  • the crude heterodimer sample was loaded on a Poly CAT A cation exchange column at 20ml/min.
  • the column was previously equilibrated with 60mM MOPS, 1 mM Calcium Acetate (CaAc) pH 6.4, at 4°C, (Buffer A).
  • Die column was washed with 150ml of Buffer A at 15ml/min.
  • a 50min linear gradient was performed at 15ml/min using Buffer A and Buffer B (60mM MOPS, 20mM CaAc pH 7.5 at 4°C).
  • Buffer C comprises 60mM MOPS, lOOmM CaCl 2 , pH 7.5.
  • Fractions 3 through 7 were pooled (total volume - 218ml), concentrated to 50ml and dialyzed against 4 liters of 60mM MOPS, 0.5mM CaAc pH 6.4 at 4°C overnight. The dialyzed pool was filtered through a O.22 ⁇ filter and checked for absorbance at 280nm. The filtrate was loaded onto the PolyCAT A column, equilibrated with 60mM MOPS, 1 M CaAc pH 6.4 at 4°C, at a flow rate of lOml/min.
  • Buffer B was changed to 60mM MOPS, lOmM CaAc pH 7.5 at 4°C.
  • the gradient was run as in Table 5.
  • the fractions were collected by peak and analyzed for activity, purity, and molecular weight.
  • the chromatogram is shown in Figure 5. Fraction identification and analysis is presented in Table 6.
  • Fractions 2, 5, and 6 correspond to the three main peaks in Figure 5 and therefore were chosen to be analyzed by HPLC size exclusion.
  • Fraction 2 corresponds to the peak that runs at 21.775 minutes in the preparative purification (Figure 5), and runs on the HPLC sizing column at 20.525 minutes, which is in the monomeric position ( Figure 7).
  • Fractions 5 and 6 (30.1 and 33.455 minutes, respectively, in Figure 5) run on the HPLC sizing column ( Figures 8 and 9) at 19.133 and 19.163 minutes, respectively (see Table 6). Therefore, both of these peaks could be considered dimers.
  • 40% Quenching assays were performed on all fractions of this purification. Only fraction 5 gave significant activity. 2.4 mg of active CC49/4-4-20 heterodimer
  • the active heterodimer Fv faction should contain both polypeptide chains. Internal sequence analysis showed that fractions 5 and 6 displayed
  • the fluorescein association constants (Ka) were determined for fractions 5 and 6 using the fluorescence quenching assay described by Herron, J.N., in Fluorescence Hapten: An Immunological Probe, E.W. Voss, ed., CRC Press, Boca Raton, FL (1984). Each sample was diluted to approximately 5.0 x lO "8 M with 20 mM HEPES buffer pH 8.0. 590 ⁇ l of the
  • the CC49 monoclonal antibody was developed by Dr. Jeffrey Schlom's group, Laboratory of Tumor Immunology and Biology, National Cancer Institute. It binds specifically to the pan-carcinoma tumor antigen TAG-72.
  • a competition enzyme-linked immunosorbent assay (ELISA) was set up in which a CC49 IgG labeled with biotin was competed against unlabeled CC49/4-4-20 Fv and the CC49/212 sFv for binding to TAG-72 on a human breast carcinoma extract (see Figure 11).
  • the amount of biotin-labeled CC49 IgG was determined using avidin, biotin coupled to horse radish peroidase in a preformed complex and o-phenylene diamine dihydrochloride (OPD). The reaction was stopped after 10 min.
  • heterodimer Fv from two complementary mixed sFv's which has been shown to have the size of a dimer of the sFv's.
  • the N-terminal analysis has shown that the active heterodimer Fv contains two polypeptide chains.
  • the heterodimer Fv has been shown to be active for both fluorescein and TAG-72 binding.
  • ADDRESSEE Sterne, Kessler, Goldstein & Fox
  • Val Arg Gly Ser Pro Ala lie Asn Val Ala Val His Val Phe 1 5 10
  • GGT AAT CAA AAG AAC TAC TTG GCC TGG TAC CAG CAG AAA CCA GGG CAG 144 Gly Asn Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45
  • TCT CCT AAA CTG CTG ATT TAC TGG GCA TCC GCT AGG GAA TCT GGG GTC 192 Ser Pro Lys Leu Leu He Tyr Trp Ala Ser Ala Arg Glu Ser Gly Val 50 55 60

Abstract

Nouveau segment de liaison de peptides utile pour relier des constituants de polypeptides en un nouveau polypeptide fusionné lié. Le segment de liaison de peptides de la présente invention présente une plus grande stabilité et est moins susceptible de s'agréger que les segments de liaison de peptides connus jusqu'ici. Il peut comporter jusqu'à environ 50 acides aminés sur sa longueur et contient au moins une fois un acide aminé chargé suivi d'une proline. Lorsqu'il est utilisé pour obtenir un fragment Fv monocaténaire, le segment de liaison de peptides possède de préférence de 18 à environ 30 acides aminés sur sa longueur. Une version préférée du segment de liaison de peptides de la présente invention comporte la séquence GSTSGSGXPGSGEGSTKG (SEQ ID NO 1) dans laquelle X est un acide aminé chargé, de préférence lysine ou arginine. Des procédés de production de polypeptides fusionnés liés à l'aide du segment de liaison de peptides de la présente invention sont décrits. Des molécules d'ADN codant lesdits polypeptides fusionnés liés et des procédés de production desdits polypeptides fusionnés liés à partir de ces molécules d'ADN sont également décrits.
PCT/US1993/011138 1992-11-20 1993-11-17 Segment de liaison pour polypeptides fusionnes lies WO1994012520A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU56701/94A AU5670194A (en) 1992-11-20 1993-11-17 Linker for linked fusion polypeptides

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US98052992A 1992-11-20 1992-11-20
US07/980,529 1992-11-20
US284593A 1993-01-15 1993-01-15
US08/002,845 1993-01-15

Publications (1)

Publication Number Publication Date
WO1994012520A1 true WO1994012520A1 (fr) 1994-06-09

Family

ID=26670961

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1993/011138 WO1994012520A1 (fr) 1992-11-20 1993-11-17 Segment de liaison pour polypeptides fusionnes lies

Country Status (3)

Country Link
US (2) US5856456A (fr)
AU (1) AU5670194A (fr)
WO (1) WO1994012520A1 (fr)

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996040915A2 (fr) * 1995-06-07 1996-12-19 Dana-Farber Cancer Institute Nouveaux ligands de ctla4/cd28 et leurs utilisations
EP0839831A2 (fr) * 1994-02-18 1998-05-06 Sensi-Test Gonadotropin monochaine
WO1998018934A1 (fr) * 1996-10-29 1998-05-07 Oxford Biomedica (Uk) Limited Gene therapeutique
GB2333527A (en) * 1996-10-29 1999-07-28 Oxford Biomedica Ltd Therapeutic gene
EP0979102A1 (fr) * 1997-04-30 2000-02-16 Enzon, Inc. Polypeptides a chaine unique modifies par oxyde de polyalkylene
US6103521A (en) * 1995-02-06 2000-08-15 Cell Genesys, Inc. Multispecific chimeric receptors
US6132722A (en) * 1997-05-07 2000-10-17 Bristol-Myers Squibb Company Recombinant antibody-enzyme fusion proteins
WO2001023543A1 (fr) * 1999-09-24 2001-04-05 Large Scale Biology Corporation Creation de regions de liaison a sequence et longueur variables, pour molecules a domaine double ou multiple
US6333396B1 (en) 1998-10-20 2001-12-25 Enzon, Inc. Method for targeted delivery of nucleic acids
US6824779B1 (en) 1993-07-26 2004-11-30 Dana-Farber Cancer Institute, Inc. Methods for inhibiting the interaction of B7-2 with its natural ligand
US6924123B2 (en) 1996-10-29 2005-08-02 Oxford Biomedica (Uk) Limited Lentiviral LTR-deleted vector
EP1801121A1 (fr) * 2005-12-23 2007-06-27 CONARIS research institute AG Variantes solubles de la molécule gp130 utiles comme médicament
US7297478B1 (en) 2000-09-22 2007-11-20 Large Scale Biology Corporation Creation of variable length and sequence linker regions for dual-domain or multi-domain molecules
US8895012B2 (en) 2006-06-30 2014-11-25 Conaris Research Institute Ag sgp130/Fc dimers
WO2015165480A1 (fr) 2014-04-30 2015-11-05 Institute For Research In Biomedicine Compositions de vaccin de cytomégalovirus humain et leur procédé de production
WO2016033168A1 (fr) * 2014-08-26 2016-03-03 University Of Washington Through Its Center For Commercialization Polypeptides et leur utilisation pour le traitement de la grippe
US9624272B2 (en) 2013-03-14 2017-04-18 University Of Washington Through Its Center For Commercialization Polypeptides for treating and/or limiting influenza infection
WO2018013563A1 (fr) * 2016-07-12 2018-01-18 Kite Pharma, Inc. Molécules de liaison à l'antigène et procédés d'utilisation associés
WO2018042385A3 (fr) * 2016-09-02 2018-04-19 The Regents Of The University Of California Méthodes et compositions impliquant des fragments variables à chaîne unique (scfv) de liaison au récepteur alpha de l'interleukine-6
US10519218B2 (en) 2014-12-01 2019-12-31 Ferring B.V. Selective IL-6-trans-signalling inhibitor compositions
JP2020534030A (ja) * 2017-09-19 2020-11-26 パウル・シェラー・インスティトゥート トランスグルタミナーゼコンジュゲーション方法およびリンカー
WO2021108482A1 (fr) 2019-11-27 2021-06-03 The Board Of Trustees Of The University Of Illinois Pentapeptide et ses méthodes d'utilisation
WO2021236879A1 (fr) 2020-05-20 2021-11-25 The Board Of Trustees Of The University Of Illinois Méthode de traitement de maladies lysosomales à l'aide de peptides d'histatine
US11198721B2 (en) 2014-12-01 2021-12-14 Ferring B.V. Methods for treatment of inflammatory disease or IL-6-mediated condition with gp130 protein
US11384350B2 (en) 2014-12-15 2022-07-12 The Regents Of The University Of California Cytotoxic molecules responsive to intracellular ligands for selective T cell mediated killing
WO2023086787A1 (fr) * 2021-11-09 2023-05-19 Janssen Biotech, Inc. Dispositif et système de co-encapsulation microfluidique et procédés d'identification de ligands de récepteur de lymphocytes t

Families Citing this family (230)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6025165A (en) * 1991-11-25 2000-02-15 Enzon, Inc. Methods for producing multivalent antigen-binding proteins
GB9225453D0 (en) * 1992-12-04 1993-01-27 Medical Res Council Binding proteins
US20060078561A1 (en) * 1994-01-31 2006-04-13 The Trustees Of Boston University Polyclonal antibody libraries
EP1231268B1 (fr) 1994-01-31 2005-07-27 Trustees Of Boston University Banques d'anticorps polyclonaux
US6136311A (en) 1996-05-06 2000-10-24 Cornell Research Foundation, Inc. Treatment and diagnosis of cancer
US20040009166A1 (en) * 1997-04-30 2004-01-15 Filpula David R. Single chain antigen-binding polypeptides for polymer conjugation
WO1999027964A1 (fr) * 1997-12-01 1999-06-10 Cfy Biomedicals, Inc. Anticorps de recombinaison multivalents destines au traitement d'infections dues au rhinovirus humain (hrv)
DE19819846B4 (de) 1998-05-05 2016-11-24 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Multivalente Antikörper-Konstrukte
US20030035798A1 (en) * 2000-08-16 2003-02-20 Fang Fang Humanized antibodies
US20030099629A1 (en) * 1999-03-11 2003-05-29 Immunomedics, Inc. Recombinant onconase and chemical conjugates and fusion proteins of recombinant onconase
US7115262B1 (en) 1999-03-16 2006-10-03 The United States Of America As Represented By The Department Of Health And Human Services Chimeric protein for prevention and treatment of HIV infection
US6432409B1 (en) * 1999-09-14 2002-08-13 Antigen Express, Inc. Hybrid peptides modulate the immune response
US20030235594A1 (en) * 1999-09-14 2003-12-25 Antigen Express, Inc. Ii-Key/antigenic epitope hybrid peptide vaccines
US9289487B2 (en) * 1999-09-14 2016-03-22 Antigen Express, Inc. II-key/antigenic epitope hybrid peptide vaccines
US20030185796A1 (en) * 2000-03-24 2003-10-02 Chiron Corporation Methods of therapy for non-hodgkin's lymphoma
EP1935431A3 (fr) 2000-05-15 2008-08-13 Health Research, Inc. Traitements anti-cancéreux utilisant une combinaison d'anticorps contre le HER2 et l'interleukine-2
US7288390B2 (en) 2000-08-07 2007-10-30 Centocor, Inc. Anti-dual integrin antibodies, compositions, methods and uses
US6902734B2 (en) 2000-08-07 2005-06-07 Centocor, Inc. Anti-IL-12 antibodies and compositions thereof
UA81743C2 (uk) 2000-08-07 2008-02-11 Центокор, Инк. МОНОКЛОНАЛЬНЕ АНТИТІЛО ЛЮДИНИ, ЩО СПЕЦИФІЧНО ЗВ'ЯЗУЄТЬСЯ З ФАКТОРОМ НЕКРОЗУ ПУХЛИН АЛЬФА (ФНПα), ФАРМАЦЕВТИЧНА КОМПОЗИЦІЯ, ЩО ЙОГО МІСТИТЬ, ТА СПОСІБ ЛІКУВАННЯ РЕВМАТОЇДНОГО АРТРИТУ
US20040072315A1 (en) * 2001-02-05 2004-04-15 Xinjie Lu Integrin-binding chimeras
JP4369662B2 (ja) 2001-04-26 2009-11-25 アビディア インコーポレイテッド 単量体ドメインのコンビナトリアルライブラリー
ATE405582T1 (de) * 2001-06-05 2008-09-15 Altor Bioscience Corp P53 bindende t-zellrezeptormoleküle und deren verwendungen
WO2003062370A2 (fr) 2001-07-19 2003-07-31 Perlan Therapeutics, Inc. Proteines multimeres et methodes de production et d'utilisation de ces proteines
US7332585B2 (en) 2002-04-05 2008-02-19 The Regents Of The California University Bispecific single chain Fv antibody molecules and methods of use thereof
US7179645B2 (en) * 2002-09-24 2007-02-20 Antigen Express, Inc. Ii-Key/antigenic epitope hybrid peptide vaccines
CA2499123A1 (fr) 2002-09-17 2004-04-15 Antigen Express, Inc. Vaccins a base d'epitope antigenique ii-key peptidique hybride
US9701754B1 (en) 2002-10-23 2017-07-11 City Of Hope Covalent disulfide-linked diabodies and uses thereof
US9708410B2 (en) 2003-05-30 2017-07-18 Janssen Biotech, Inc. Anti-tissue factor antibodies and compositions
ATE447611T1 (de) 2003-08-18 2009-11-15 Univ California Polypeptid-display-bibliotheken und verfahren zur herstellung und verwendung davon
WO2005040382A2 (fr) * 2003-10-28 2005-05-06 Alexander Cherkasky Proteines de fusion de cherkasky contenant des zones de liaison a des anticorps, a des antigenes et a des microtubules, et des zones declenchant une reponse immunitaire
US7304139B2 (en) * 2003-10-28 2007-12-04 University Of Florida Research Foundation, Inc. Polynucleotides and polypeptides of Anaplasma phagocytophilum and methods of using the same
DK1694360T3 (da) 2003-11-04 2010-10-18 Novartis Vaccines & Diagnostic Anvendelse af antagonist anti-CD40-antistoffer til behandling af autoimmune og inflammatoriske sygdomme og organtransplantationafstødning
AU2004287480B2 (en) 2003-11-04 2011-09-15 Novartis Vaccines And Diagnostics, Inc. Use of antagonist anti-CD40 antibodies for treatment of chronic lymphocytic leukemia
PL1684805T3 (pl) 2003-11-04 2010-12-31 Novartis Vaccines & Diagnostics Inc Sposoby leczenia szpiczaka mnogiego z zastosowaniem antagonistycznych monoklonalnych przeciwciał przeciwko CD40
ATE476991T1 (de) 2003-11-04 2010-08-15 Novartis Vaccines & Diagnostic Verfahren zur behandlung von soliden tumoren mit expression des cd40-zelloberflächen-antigens
CA2544368C (fr) 2003-11-04 2014-04-01 Chiron Corporation Procedes de therapie pour des cancers lies a la cellule b
US7576183B2 (en) * 2003-12-24 2009-08-18 Los Alamos National Security, Llc Structure-based receptor MIMICS targeted against bacterial superantigen toxins
US7595380B2 (en) 2005-04-27 2009-09-29 Tripath Imaging, Inc. Monoclonal antibodies and methods for their use in the detection of cervical disease
PE20061324A1 (es) 2005-04-29 2007-01-15 Centocor Inc Anticuerpos anti-il-6, composiciones, metodos y usos
ES2429564T3 (es) 2005-05-18 2013-11-15 Novartis Ag Procedimientos para el diagnóstico y tratamiento de enfermedades que tienen un componente autoinmune y/o inflamatorio
EP2452694B1 (fr) 2005-06-30 2018-11-14 Janssen Biotech, Inc. Anticorps anti-IL-23, compositions, procédés et utilisations
US20070026012A1 (en) * 2005-08-01 2007-02-01 Cornell Research Foundation, Inc. Compositions and methods for monitoring and altering protein folding and solubility
EP1917347B1 (fr) * 2005-08-22 2015-10-07 Cornell Research Foundation, Inc. Compositions et methodes permettant d'analyser des interactions de proteines
AU2006284651C1 (en) 2005-08-31 2013-09-12 The Regents Of The University Of California Cellular libraries of peptide sequences (CLiPS) and methods of using the same
US7632498B2 (en) 2005-12-19 2009-12-15 Tripath Imaging, Inc. MCM6 and MCM7 monoclonal antibodies and methods for their use in the detection of cervical disease
PT3219328T (pt) 2005-12-29 2020-08-28 Janssen Biotech Inc Anticorpos anti-il-23 humana, composições, método e usos
GEP20135917B (en) 2006-03-17 2013-09-10 Biogen Idec Inc Stabilized polypeptide compositions
CA2646329C (fr) 2006-03-20 2018-07-03 The Regents Of The University Of California Anticorps modifies diriges contre l'antigene de cellules souches prostatiques servant a cibler le cancer
ES2395969T3 (es) 2006-03-24 2013-02-18 Merck Patent Gmbh Dominios de proteínas heterodiméricas genéticamente modificados
US7612178B2 (en) * 2006-03-28 2009-11-03 Biogen Idec Ma Inc Anti-IGF-1R antibodies and uses thereof
US8945564B2 (en) 2006-04-21 2015-02-03 Novartis Ag Antagonist anti-CD40 antibody pharmaceutical compositions
US7727525B2 (en) * 2006-05-11 2010-06-01 City Of Hope Engineered anti-CD20 antibody fragments for in vivo targeting and therapeutics
EP1878744A1 (fr) * 2006-07-13 2008-01-16 Max-Delbrück-Centrum für Molekulare Medizin (MDC) Epitope-tag pour des récepteurs de cellules T exprimés en surface, leurs utilisations et méthode de selection de cellules hôtes les exprimant
US20080035574A1 (en) * 2006-08-08 2008-02-14 Sabottke Craig Y Membrane Barrier films and method of use
US8580263B2 (en) * 2006-11-21 2013-11-12 The Regents Of The University Of California Anti-EGFR family antibodies, bispecific anti-EGFR family antibodies and methods of use thereof
AU2007353773A1 (en) * 2006-12-22 2008-11-27 Dow Agrosciences Llc Plant-made West Nile virus (WNV) vaccines, vectors and plant codon optimized sequences
WO2008089132A2 (fr) * 2007-01-12 2008-07-24 Cornell Research Foundation, Inc. Sélection génétique pour le repliement des protéines et la solubilité dans le périplasme bactérien
WO2008118324A2 (fr) 2007-03-26 2008-10-02 Macrogenics, Inc. Composition et procédé de traitement du cancer avec un anticorps anti-uroplakine ib
ES2614735T3 (es) 2007-07-23 2017-06-01 Janssen Biotech, Inc. Métodos y composiciones para tratar los trastornos relacionados con fibrosis usando antagonistas de la IL-17
WO2009014726A1 (fr) 2007-07-26 2009-01-29 The Regents Of The University Of California Procédés permettant d'améliorer un affichage de cellules bactériennes de protéines et de peptides
EP2190480A4 (fr) * 2007-08-28 2013-01-23 Biogen Idec Inc Anticorps anti-igf-1r et leurs utilisations
JP2010538012A (ja) * 2007-08-28 2010-12-09 バイオジェン アイデック マサチューセッツ インコーポレイテッド Ｉｇｆ−１ｒの複数のエピトープに結合する組成物
JP6126773B2 (ja) 2007-09-04 2017-05-10 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 癌のターゲッティングおよび検出のための高親和性抗前立腺幹細胞抗原（ｐｓｃａ）抗体
JP5298134B2 (ja) 2007-11-01 2013-09-25 パーシード セラピューティクス リミテッド ライアビリティ カンパニー 免疫抑制ポリペプチドおよび核酸
MX2010008578A (es) 2008-02-08 2010-11-10 Medimmune Llc Anticuerpos anti-receptor 1 de interferon alfa con afinidad reducida al ligando fc.
EP2274008B1 (fr) 2008-03-27 2014-02-26 ZymoGenetics, Inc. Compositions et procédés pour inhiber pdgfr-bêta et vegf-a
JP2011516549A (ja) * 2008-04-11 2011-05-26 バイオジェン アイデック マサチューセッツ インコーポレイテッド 抗ｉｇｆ−１ｒ抗体と他の化合物との治療的併用
CA2733642A1 (fr) 2008-08-14 2010-02-18 Cephalon Australia Pty Ltd Anticorps anti-il-12/il-23
US8748130B2 (en) * 2008-09-02 2014-06-10 Antigen Express, Inc. Human papillomavirus / Ii-Key hybrids and methods of use
US10131693B2 (en) 2008-10-20 2018-11-20 Gwangju Institute Of Science And Technology Bipodal-peptide binder
CA3149920A1 (fr) 2008-10-31 2010-05-06 Janssen Biotech, Inc. Compositions d'echafaudage a base de domaines de la fibronectine de type iii, procedes et utilisations
US8927694B2 (en) * 2008-11-18 2015-01-06 Merrimack Pharmaceuticals, Inc. Human serum albumin linkers and conjugates thereof
US20100310591A1 (en) * 2009-01-28 2010-12-09 Robert Humphreys Ii-KEY HYBRID PEPTIDES THAT MODULATE THE IMMUNE RESPONSE TO INFLUENZA
AU2010213892B2 (en) 2009-02-12 2014-10-23 Janssen Biotech, Inc. Fibronectin type III domain based scaffold compositions, methods and uses
US10517969B2 (en) 2009-02-17 2019-12-31 Cornell University Methods and kits for diagnosis of cancer and prediction of therapeutic value
WO2010102167A1 (fr) 2009-03-05 2010-09-10 Becton, Dickinson And Company Anticorps monoclonaux de métalloprotéinase-7 matricielle (mmp-7) et méthodes d'utilisation pour la détection du cancer de l'ovaire
EP2403875A1 (fr) 2009-03-06 2012-01-11 Tripath Imaging, Inc. Anticorps monoclonaux de la glycodeline et leurs procedes d'utilisation dans la detection du cancer de l'ovaire
WO2011063198A2 (fr) 2009-11-20 2011-05-26 St. Jude Children's Research Hospital Procédés et compositions de modulation de l'activité du complexe du récepteur de l'interleukine 35
RU2673908C2 (ru) 2009-12-02 2018-12-03 Имэджинэб, Инк. Мини-антитела j591 и цис-диатела для направленной доставки простата-специфичного мембранного антигена (psma) человека и способы их применения
US20110183891A1 (en) 2009-12-16 2011-07-28 Jordan Tang Methods and Compositions for Inhibiting Fungal Infection and Disease
EP2513137B1 (fr) 2009-12-17 2018-02-28 Children's Medical Center Corporation Peptides dérivés de saposine a et utilisations de ceux-ci
CA2789629A1 (fr) 2010-02-10 2011-08-18 Immunogen, Inc. Anticorps anti-cd20 et utilisations de ceux-ci
US20130045871A1 (en) 2010-03-18 2013-02-21 Cornell University Engineering correctly folded antibodies using inner membrane display of twin-arginine translocation intermediates
WO2011115688A2 (fr) 2010-03-19 2011-09-22 H. Lee Moffitt Cancer Center & Research Institute Inhibiteurs des interactions de l'intégrine pour traiter le cancer
US8383793B2 (en) 2010-04-15 2013-02-26 St. Jude Children's Research Hospital Methods and compositions for the diagnosis and treatment of cancer resistant to anaplastic lymphoma kinase (ALK) kinase inhibitors
WO2011143274A1 (fr) 2010-05-10 2011-11-17 Perseid Therapeutics Polypeptides inhibiteurs de vla4
CA2802017A1 (fr) 2010-06-09 2011-12-15 Zymogenetics, Inc. Proteines de fusion vstm3 dimeres et compositions et procedes connexes
JP2013539453A (ja) 2010-07-09 2013-10-24 エクセリクシス， インク． 癌の治療のためのキナーゼ阻害物質の組み合わせ
WO2012075111A1 (fr) 2010-11-30 2012-06-07 Novartis Ag Utilisation d'anticorps anti-cd40 en thérapie combinée contre des cancers associés aux cellules b
MX2013007392A (es) 2010-12-22 2013-11-01 Cephalon Australia Pty Ltd Anticuerpo modificado con vida media mejorada.
CN103732628B (zh) 2011-03-03 2017-06-23 酵活有限公司 多价杂多聚体骨架设计和构建体
WO2012142233A1 (fr) 2011-04-14 2012-10-18 St. Jude Children's Research Hospital Procédés et compositions pour détecter et moduler un nouveau complexe mtor
EP3540100B8 (fr) 2011-09-27 2022-11-02 Janssen Biotech, Inc. Échafaudage de protéine à base de répétition de fibronectine de type iii avec des surfaces de liaison alternatives
AU2012358269B2 (en) 2011-12-22 2017-11-02 Children's Medical Center Corporation Saposin-A derived peptides and uses thereof
US9062120B2 (en) 2012-05-02 2015-06-23 Janssen Biotech, Inc. Binding proteins having tethered light chains
MX355041B (es) 2012-05-25 2018-03-28 Janssen Biotech Inc Dominios de union a albumina consenso no naturales.
DK2864360T3 (en) 2012-06-25 2017-12-18 Brigham & Womens Hospital Inc TARGETED THERAPY
UA114108C2 (uk) 2012-07-10 2017-04-25 Борд Оф Ріджентс, Дзе Юніверсіті Оф Техас Сістем Моноклональне антитіло для застосування в діагностиці і терапії злоякісних пухлин і аутоімунного захворювання
WO2014012082A2 (fr) 2012-07-13 2014-01-16 Zymeworks Inc. Conception d'échafaudages hétéromultimères et produits de recombinaison
KR101443839B1 (ko) * 2012-07-27 2014-09-26 광주과학기술원 펩타이드의 타겟 친화도 개선방법
WO2014036520A1 (fr) 2012-08-30 2014-03-06 Merrimack Pharmaceuticals, Inc. Polythérapies comprenant des agents anti-erbb3
EP4029937A1 (fr) 2012-10-04 2022-07-20 Research Development Foundation Molécules et thérapies de sérine protéase
LT3447069T (lt) 2012-11-21 2020-12-10 Janssen Biotech, Inc. Bispecifiniai egfr/c-met antikūnai
US9695228B2 (en) 2012-11-21 2017-07-04 Janssen Biotech, Inc. EGFR and c-Met fibronectin type III domain binding molecules
US9388397B2 (en) 2013-02-15 2016-07-12 Research Development Foundation Deimmunized gelonin molecules and therapies
WO2014134179A1 (fr) 2013-02-28 2014-09-04 The Board Of Regents Of The University Of Texas System Procédés pour classer un cancer comme sensible aux thérapies dirigées par tmepai et pour traiter ces cancers
JP6457999B2 (ja) 2013-03-14 2019-01-23 ザ チルドレンズ メディカル センター コーポレーション 処置に関してがんの対象を識別するためのｃｄ３６の使用
WO2015057545A2 (fr) 2013-10-14 2015-04-23 Janssen Biotech, Inc. Molécules de liaison au domaine de fibronectine de type iii modifiées par la cystéine
EP3074419B1 (fr) 2013-11-25 2018-08-29 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Récepteurs d'antigènes chimériques pour lutter contre une infection par le vih
KR102534681B1 (ko) 2013-12-24 2023-05-18 잔센파마슈티카엔.브이. 항-vista 항체 및 단편
JP6908381B2 (ja) 2014-01-29 2021-07-28 デイナ ファーバー キャンサー インスティチュート，インコーポレイテッド Ｍｕｃ１−ｃ／細胞外ドメイン（ｍｕｃ１−ｃ／ｅｃｄ）に対する抗体
US9738702B2 (en) 2014-03-14 2017-08-22 Janssen Biotech, Inc. Antibodies with improved half-life in ferrets
EP4154901A1 (fr) 2014-03-26 2023-03-29 The Children's Medical Center Corporation Peptides de prosaposine cycliques et leurs utilisations
KR20150118252A (ko) * 2014-04-11 2015-10-22 한국과학기술원 환형의 β-헤어핀 기반 펩타이드 바인더 및 이의 제조방법
BR112016024088A2 (pt) 2014-04-16 2018-06-19 Biocon Ltd. formulações de proteína estáveis que compreendem um excesso molar de sorbitol
WO2015179435A1 (fr) 2014-05-19 2015-11-26 Bayer Healthcare Llc Anticorps monoclonaux humanisés optimisés dirigés contre la protéine c activée et leurs utilisations
WO2016001810A1 (fr) 2014-07-01 2016-01-07 Pfizer Inc. Diabodies hétérodimères bispécifiques et leurs utilisations
EP3215611B1 (fr) 2014-11-06 2019-08-21 E. I. du Pont de Nemours and Company Administration médiée par un peptide d'endonucléase guidée par un arn dans des cellules
US10479825B2 (en) 2015-02-25 2019-11-19 Vanderbilt University Antibody-mediated neutralization of Marburg virus
US11046959B2 (en) 2015-03-30 2021-06-29 The Board Of Regents Of The Nevada System Of Higher Education On Behalf Of The University Of Nevada, Las Vegas Compositions comprising TALENs and methods of treating HIV
KR102625793B1 (ko) 2015-05-06 2024-01-15 얀센 바이오테크 인코포레이티드 전립선 특이적 막 항원 결합 피브로넥틴 iii형 도메인
US10844122B2 (en) 2015-05-06 2020-11-24 Janssen Biotech, Inc. Prostate specific membrane antigen (PSMA) bispecific binding agents and uses thereof
BR112017027870A2 (pt) 2015-06-24 2018-08-28 Janssen Pharmaceutica Nv anticorpos e fragmentos anti-vista
CA3168241A1 (fr) 2015-07-15 2017-01-19 Rutgers. The State University of New Jersey Plate-forme d'edition genique ciblee sans nuclease et utilisations de celle-ci
CN115960230A (zh) 2015-08-07 2023-04-14 伊麦吉纳博公司 靶向分子的抗原结合构建体
EP3328881B1 (fr) 2015-09-08 2019-08-28 Theripion, Inc. Polypeptides de fusion apoa-1 ainsi que compositions et procédés associés
AU2016354440B2 (en) 2015-11-09 2023-09-28 The Children's Hospital Of Philadelphia Glypican 2 as a cancer marker and therapeutic target
BR112018016461A2 (pt) 2016-02-12 2019-10-01 Janssen Pharmaceutica Nv anticorpos e fragmentos anti-vista, usos dos mesmos e métodos de identificação dos mesmos
EP3417058B1 (fr) 2016-02-16 2021-09-22 Research Development Foundation Molécules modifiées par sortase et utilisations de celles-ci
CA3019164A1 (fr) 2016-03-29 2017-10-05 Janssen Biotech, Inc. Methode de traitement du psoriasis par l'augmentation de l'espacement des doses d'anticorps anti-il-12/23
WO2017175058A1 (fr) 2016-04-07 2017-10-12 Janssen Pharmaceutica Nv Anticorps anti-vista et fragments de ceux-ci, leurs utilisations et leurs procédés d'identification
EA201892796A1 (ru) 2016-06-03 2019-06-28 Янссен Байотек, Инк. Связывающиеся с сывороточным альбумином домены фибронектина типа iii
AU2017281083B2 (en) 2016-06-21 2022-01-27 Janssen Biotech, Inc. Cysteine engineered fibronectin type III domain binding molecules
WO2018014039A1 (fr) 2016-07-15 2018-01-18 Poseida Therapeutics, Inc. Récepteurs d'antigènes chimériques (cars) spécifiques pour muc1 et leurs procédés d'utilisation
MX2019000643A (es) 2016-07-15 2019-06-13 Poseida Therapeutics Inc Receptores de antigenos quimericos y metodos de uso.
US9963498B2 (en) 2016-08-18 2018-05-08 Wisconsin Alumni Research Foundation Peptides that inhibit syndecan-1 activation of VLA-4 and IGF-1R
CA3037961A1 (fr) 2016-09-30 2018-04-05 Janssen Biotech, Inc. Procede sur et efficace de traitement du psoriasis avec un anticorps specifique contre l'il-23
MX2019005661A (es) 2016-11-16 2019-10-07 Janssen Biotech Inc Método para tratar la psoriasis con el anticuerpo específico anti-il23.
WO2018106993A1 (fr) * 2016-12-09 2018-06-14 H. Lee Moffitt Cancer Center And Research Institute Inc. Récepteurs antigéniques chimériques se liant à tlr9
WO2018136163A2 (fr) 2016-12-09 2018-07-26 Theripion, Inc. Polypeptides de fusion apoa-1 en tandem
WO2018111976A1 (fr) 2016-12-14 2018-06-21 Janssen Biotech, Inc. Domaines de fibronectine de type iii se liant à pd-l1
US10626165B2 (en) 2016-12-14 2020-04-21 Janssen Biotech, Inc. CD8a-binding fibronectin type III domains
WO2018111978A1 (fr) 2016-12-14 2018-06-21 Janssen Biotech, Inc. Domaines de fibronectine de type iii à liaison au cd137
US11519009B2 (en) 2017-01-09 2022-12-06 University Of Massachusetts Complexes for gene deletion and editing
CN110234351A (zh) 2017-01-30 2019-09-13 詹森生物科技公司 用于治疗活动性银屑病关节炎的抗tnf抗体、组合物和方法
MX2019009377A (es) 2017-02-07 2019-12-11 Janssen Biotech Inc Anticuerpos anti-tnf, composiciones y metodos para el tratamiento de la espondilitis anquilosante activa.
US11266745B2 (en) 2017-02-08 2022-03-08 Imaginab, Inc. Extension sequences for diabodies
US11744861B2 (en) 2017-03-13 2023-09-05 Poseida Therapeutics, Inc. Compositions and methods for selective elimination and replacement of hematopoietic stem cells
CN108690138A (zh) * 2017-04-12 2018-10-23 鸿运华宁（杭州）生物医药有限公司 一种能与人cd19或cd20和人cd3结合的双特异性抗体及其应用
JP7250766B2 (ja) 2017-08-25 2023-04-03 ヤンセン バイオテツク，インコーポレーテツド ＦＣγＲＩＩ結合フィブロネクチンＩＩＩ型ドメイン、それらのコンジュゲート、及びそれらを含む多重特異性分子
EP3679145A2 (fr) 2017-09-08 2020-07-15 Poseida Therapeutics, Inc. Compositions et procédés d'expression génique conditionnelle médiée par un récepteur de ligand chimérique (clr)
TW201922780A (zh) 2017-09-25 2019-06-16 美商健生生物科技公司 以抗ｉｌ１２／ｉｌ２３抗體治療狼瘡之安全且有效之方法
US11198722B2 (en) 2017-10-06 2021-12-14 University Of Utah Research Foundation Immune tolerant elastin-like peptide tetramer guided nanoparticles and methods of use
WO2019075097A1 (fr) 2017-10-11 2019-04-18 Board Of Regents, The University Of Texas System Anticorps pd-l1 humains et leurs procédés d'utilisation
WO2019156758A2 (fr) 2018-01-05 2019-08-15 Vanderbilt University Neutralisation à médiation par anticorps du virus chikungunya
WO2019150309A1 (fr) 2018-02-02 2019-08-08 Hammack Scott Modulateurs de gpr68 et leurs utilisations pour le traitement et la prévention de maladies
EP3762015A4 (fr) 2018-03-05 2022-04-27 Janssen Biotech, Inc. Méthodes de traitement de la maladie de crohn avec un anticorps spécifique anti-il23
US20210214445A1 (en) 2018-03-23 2021-07-15 Board Of Regents, The University Of Texas System Dual specificity antibodies to pd-l1 and pd-l2 and methods of use therefor
EP3790587A4 (fr) 2018-05-11 2022-01-26 Janssen Biotech, Inc. Méthodes de traitement de la dépression à l'aide d'anticorps il-23
WO2019241315A1 (fr) 2018-06-12 2019-12-19 Obsidian Therapeutics, Inc. Constructions régulatrices dérivées de pde5 et procédés d'utilisation en immunothérapie
US20210301305A1 (en) 2018-06-13 2021-09-30 Voyager Therapeutics, Inc. Engineered untranslated regions (utr) for aav production
CN108707198A (zh) * 2018-06-28 2018-10-26 李永海 识别人c-Met蛋白的人源单链抗体、诊断试剂及其CAR-T细胞制剂
WO2020016838A2 (fr) 2018-07-18 2020-01-23 Janssen Biotech, Inc. Prédicteurs de réponse prolongée après le traitement avec un anticorps spécifique anti-il23
WO2020041360A1 (fr) 2018-08-21 2020-02-27 Quidel Corporation Anticorps dbpa et leurs utilisations
CN112672760A (zh) 2018-09-13 2021-04-16 德克萨斯大学体系董事会 新颖lilrb4抗体和其用途
FI3883606T3 (fi) 2018-09-24 2023-09-07 Janssen Biotech Inc Turvallinen ja tehokas menetelmä haavaisen paksusuolitulehduksen hoitamiseksi anti-il12/il23-vasta-aineella
CA3115248A1 (fr) 2018-10-05 2020-04-09 Voyager Therapeutics, Inc. Constructions d'acides nucleiques modifies codant pour des proteines de production d'aav
US20210386788A1 (en) 2018-10-24 2021-12-16 Obsidian Therapeutics, Inc. Er tunable protein regulation
US11548941B2 (en) 2018-11-20 2023-01-10 Janssen Biotech, Inc. Safe and effective method of treating psoriasis with anti-IL-23 specific antibody
US20200197517A1 (en) 2018-12-18 2020-06-25 Janssen Biotech, Inc. Safe and Effective Method of Treating Lupus with Anti-IL12/IL23 Antibody
JP2022513507A (ja) 2018-12-20 2022-02-08 ポセイダ セラピューティクス，インコーポレイティド ナノトランスポゾン組成物および使用方法
EP3911746A1 (fr) 2019-01-14 2021-11-24 Institut National de la Santé et de la Recherche Médicale (INSERM) Procédés et kits de génération et de sélection de variante de protéine de liaison avec une affinité et/ou une spécificité de liaison accrues
AU2020208828A1 (en) 2019-01-15 2021-08-05 Janssen Biotech, Inc. Anti-TNF antibody compositions and methods for the treatment of juvenile idiopathic arthritis
EP3911410A1 (fr) 2019-01-18 2021-11-24 Voyager Therapeutics, Inc. Procédés et systèmes de fabrication de particules aav
CA3127748A1 (fr) 2019-01-23 2020-07-30 Janssen Biotech, Inc. Compositions d'anticorps anti-tnf destinees a etre utilisees dans des methodes de traitement d'arthrite psoriasique
CA3133388A1 (fr) 2019-03-14 2020-09-17 Janssen Biotech, Inc. Procedes de production de compositions d'anticorps anti-tnf
US20220144934A1 (en) 2019-03-14 2022-05-12 Janssen Biotech, Inc. Methods for Producing Anti-TNF Antibody Compositions
WO2020183269A1 (fr) 2019-03-14 2020-09-17 Janssen Biotech, Inc. Procédés de fabrication pour la production de compositions d'anticorps anti-tnf
CN113825765A (zh) 2019-03-14 2021-12-21 詹森生物科技公司 用于制备抗il12/il23抗体组合物的制造方法
EA202192459A1 (ru) 2019-03-18 2021-11-25 Янссен Байотек, Инк. Способ лечения псориаза антителом к il12/il23 у субъектов детского возраста
SG11202110626SA (en) 2019-03-27 2021-10-28 Umc Utrecht Holding Bv Engineered iga antibodies and methods of use
EP3966227A1 (fr) 2019-05-07 2022-03-16 Voyager Therapeutics, Inc. Compositions et méthodes d'augmentation vectorisée de la destruction, de l'expression et/ou de la régulation de protéines
CA3138241A1 (fr) 2019-05-23 2020-11-26 Janssen Biotech, Inc. Methode de traitement de maladie intestinale inflammatoire au moyen d'une polytherapie d'anticorps diriges contre il -23 et tnf alpha
MX2021014885A (es) 2019-06-03 2022-04-06 Janssen Biotech Inc Composiciones de anticuerpos anti-tnf, y métodos para el tratamiento de la artritis psoriásica.
JP2022535534A (ja) 2019-06-03 2022-08-09 ヤンセン バイオテツク，インコーポレーテツド 活動性強直性脊椎炎を治療するための抗ｔｎｆ抗体、組成物、及び方法
JP2022541538A (ja) 2019-07-19 2022-09-26 ザ・チルドレンズ・ホスピタル・オブ・フィラデルフィア グリピカン２結合ドメインを含有するキメラ抗原受容体
EP4004037A1 (fr) 2019-07-26 2022-06-01 Vanderbilt University Anticorps monoclonaux humains dirigés contre l'entérovirus d68
WO2021028752A1 (fr) 2019-08-15 2021-02-18 Janssen Biotech, Inc. Anticorps anti-tfn pour le traitement du diabète de type i
JP2022546675A (ja) 2019-08-19 2022-11-07 ウニベルシテート バーゼル 細胞治療の方法
WO2021046155A1 (fr) 2019-09-03 2021-03-11 Voyager Therapeutics, Inc. Édition vectorisée d'acides nucléiques pour corriger des mutations manifestes
CN114761424A (zh) 2019-09-05 2022-07-15 波赛达治疗公司 同种异体细胞组合物和使用方法
US20220348937A1 (en) 2019-09-06 2022-11-03 Obsidian Therapeutics, Inc. Compositions and methods for dhfr tunable protein regulation
WO2021076546A1 (fr) 2019-10-14 2021-04-22 Aro Biotherapeutics Company Domaines de type iii de fibronectine de liaison à cd137
WO2021076574A2 (fr) 2019-10-14 2021-04-22 Aro Biotherapeutics Company Conjugués domaine fn3-arnsi et leurs utilisations
EP4077376A2 (fr) 2019-12-19 2022-10-26 Quidel Corporation Fusion d'anticorps monoclonaux
CN115135672A (zh) 2019-12-20 2022-09-30 波赛达治疗公司 抗muc1组合物和使用方法
KR20220140568A (ko) 2020-02-11 2022-10-18 벤더르빌트 유니버시티 중증 급성 호흡기 증후군 코로나바이러스 2(SARS-CoV-2)에 대한 인간 단일클론 항체
WO2021183795A1 (fr) 2020-03-11 2021-09-16 Poseida Therapeutics, Inc. Récepteurs stimulateurs chimériques et procédés d'utilisation dans l'activation et la différenciation de lymphocytes t
CA3175110A1 (fr) 2020-03-26 2021-09-30 James E. Crowe, Jr. Anticorps monoclonaux humains diriges contre le coronavirus 2 du syndrome respiratoire aigu severe (sars-cov-2)
WO2021195385A1 (fr) 2020-03-26 2021-09-30 Vanderbilt University Anticorps monoclonaux humains dirigés contre le coronavirus du syndrome respiratoire aigu sévère 2 (sras-cov-2)
MX2022012956A (es) 2020-04-14 2023-03-27 Poseida Therapeutics Inc Composiciones y métodos para su uso en el tratamiento del cáncer.
CN116249712A (zh) 2020-04-15 2023-06-09 沃雅戈治疗公司 Tau结合化合物
EP4179079A1 (fr) 2020-07-10 2023-05-17 Horizon Discovery Limited Procédé de production de cellules génétiquement modifiées
IL299928A (en) 2020-07-27 2023-03-01 Voyager Therapeutics Inc Preparations and methods for treating neurological disorders associated with glucosylcremidase beta deficiency
US20240000969A1 (en) 2020-10-21 2024-01-04 Poseida Therapeutics San Diego Compositions and methods for delivery of nucleic acids
GB202105804D0 (en) 2020-11-20 2021-06-09 Univ Cape Town Use of microvirin in the identification of mycobacterium tuberculosis mannose-capped lipoarabinomannan
US20240067954A1 (en) 2021-01-05 2024-02-29 Horizon Discovery Limited Method for producing genetically modified cells
WO2022178078A1 (fr) 2021-02-19 2022-08-25 Theripion, Inc. Polypeptides de fusion paraoxonase et compositions et méthodes associées
IL305802A (en) 2021-03-12 2023-11-01 Janssen Biotech Inc A safe and effective method for the treatment of rheumatoid arthritis with a specific anti-IL23 antibody
BR112023018400A2 (pt) 2021-03-12 2023-12-12 Janssen Biotech Inc Método para tratamento de pacientes de artrite psoriática com resposta inadequada à terapia de tnf com anticorpo específico anti-il23
IL309987A (en) 2021-07-09 2024-03-01 Janssen Biotech Inc Production methods for the production of anti-IL12/IL23 antibody compositions
AU2022306144A1 (en) 2021-07-09 2024-02-22 Janssen Biotech, Inc. Manufacturing methods for producing anti-tnf antibody compositions
AU2022308201A1 (en) 2021-07-09 2024-02-22 Janssen Biotech, Inc. Manufacturing methods for producing anti-tnf antibody compositions
WO2023023055A1 (fr) 2021-08-16 2023-02-23 Renagade Therapeutics Management Inc. Compositions et procédés d'optimisation du tropisme de systèmes d'administration d'arn
CA3231523A1 (fr) 2021-09-14 2023-03-23 Renagade Therapeutics Management Inc. Lipides acycliques et leurs procedes d'utilisation
TW202328067A (zh) 2021-09-14 2023-07-16 美商雷納嘉德醫療管理公司 環狀脂質及其使用方法
WO2023044483A2 (fr) 2021-09-20 2023-03-23 Voyager Therapeutics, Inc. Compositions et procédés pour le traitement du cancer positif her2
WO2023060088A1 (fr) 2021-10-04 2023-04-13 Poseida Therapeutics, Inc. Compositions de transposons et leurs procédés d'utilisation
WO2023073615A1 (fr) 2021-10-29 2023-05-04 Janssen Biotech, Inc. Méthodes de traitement de la maladie de crohn avec un anticorps spécifique anti-il23
WO2023084488A1 (fr) 2021-11-15 2023-05-19 Janssen Biotech, Inc. Méthodes de traitement de la maladie de crohn au moyen d'un anticorps spécifique anti-il23
WO2023091949A2 (fr) 2021-11-17 2023-05-25 Voyager Therapeutics, Inc. Compositions et méthodes de traitement de troubles neurologiques liés à un déficit en bêta glucosylcéramidase
US20230159633A1 (en) 2021-11-23 2023-05-25 Janssen Biotech, Inc. Method of Treating Ulcerative Colitis with Anti-IL23 Specific Antibody
WO2023122752A1 (fr) 2021-12-23 2023-06-29 Renagade Therapeutics Management Inc. Lipides contraints et procédés d'utilisation associés
WO2023141576A1 (fr) 2022-01-21 2023-07-27 Poseida Therapeutics, Inc. Compositions et procédés d'administration d'acides nucléiques
US20230312703A1 (en) 2022-03-30 2023-10-05 Janssen Biotech, Inc. Method of Treating Psoriasis with IL-23 Specific Antibody
US20230365708A1 (en) 2022-04-01 2023-11-16 Board Of Regents, The University Of Texas System Dual specificity antibodies to human pd-l1 and pd-l2 and methods of use therefor
GB202204813D0 (en) 2022-04-01 2022-05-18 Bradcode Ltd Human monoclonal antibodies and methods of use thereof
WO2023196931A1 (fr) 2022-04-07 2023-10-12 Renagade Therapeutics Management Inc. Lipides cycliques et nanoparticules lipidiques (npl) pour l'apport d'acides nucléiques ou de peptides destinés à être utilisés dans la vaccination contre des agents infectieux
US20230374122A1 (en) 2022-05-18 2023-11-23 Janssen Biotech, Inc. Method for Evaluating and Treating Psoriatic Arthritis with IL23 Antibody
WO2023250388A1 (fr) 2022-06-22 2023-12-28 Voyager Therapeutics, Inc. Composés se liant à la protéine tau

Family Cites Families (40)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2286443A1 (fr) * 1974-09-16 1976-04-23 Anvar Appareil pour le codage et la visualisation simultanes d'un graphe
FR2426292A1 (fr) * 1978-05-18 1979-12-14 Thomson Csf Processeur pour terminal graphique
US4414629A (en) * 1979-04-02 1983-11-08 Waite John H Method and apparatus for making correlations and predictions using a finite field of data of unorganized and/or partially structured elements
US4355023A (en) * 1980-09-30 1982-10-19 The Massachusetts General Hospital Antibody fragment compositions and process
US4434156A (en) * 1981-10-26 1984-02-28 The Salk Institute For Biological Studies Monoclonal antibodies specific for the human transferrin receptor glycoprotein
US4444878A (en) * 1981-12-21 1984-04-24 Boston Biomedical Research Institute, Inc. Bispecific antibody determinants
DE3382317D1 (de) * 1982-03-15 1991-07-25 Schering Corp Hybride dns, damit hergestellte bindungszusammensetzung und verfahren dafuer.
US4470925A (en) * 1982-05-05 1984-09-11 E. I. Du Pont De Nemours And Company Immunoglobulin half-molecules and process for producing hybrid antibodies
US4479895A (en) * 1982-05-05 1984-10-30 E. I. Du Pont De Nemours And Company Immunoglobulin half-molecules and process for producing hybrid antibodies
GB8308235D0 (en) * 1983-03-25 1983-05-05 Celltech Ltd Polypeptides
US4816567A (en) * 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
GB8422238D0 (en) * 1984-09-03 1984-10-10 Neuberger M S Chimeric proteins
US5160723A (en) * 1985-04-19 1992-11-03 Sloan-Kettering Institute For Cancer Research Method of imaging colorectal carcinoma lesion and composition for use therein
US4908773A (en) * 1987-04-06 1990-03-13 Genex Corporation Computer designed stabilized proteins and method for producing same
US4946778A (en) * 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
WO1988001649A1 (fr) * 1986-09-02 1988-03-10 Genex Corporation Molecules de liaison de chaines de polypeptide simples
US4881175A (en) * 1986-09-02 1989-11-14 Genex Corporation Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides
US4704692A (en) * 1986-09-02 1987-11-03 Ladner Robert C Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides
US5260203A (en) * 1986-09-02 1993-11-09 Enzon, Inc. Single polypeptide chain binding molecules
EP0305500B1 (fr) * 1987-03-20 1994-11-09 Creative Biomolecules, Inc. Procede de purification de polypeptides recombinants
EP0305481B1 (fr) * 1987-03-20 1993-09-29 Creative Biomolecules, Inc. Sequences d'amorce pour la production de proteines recombinantes
US4853871A (en) * 1987-04-06 1989-08-01 Genex Corporation Computer-based method for designing stablized proteins
AU612370B2 (en) * 1987-05-21 1991-07-11 Micromet Ag Targeted multifunctional proteins
US5132405A (en) * 1987-05-21 1992-07-21 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5091513A (en) * 1987-05-21 1992-02-25 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5258498A (en) * 1987-05-21 1993-11-02 Creative Biomolecules, Inc. Polypeptide linkers for production of biosynthetic proteins
DE3853740T2 (de) * 1987-06-10 1995-11-09 Dana Farber Cancer Inst Inc Bifunktionelle Antikörperkonstruktionen und Verfahren zur selektiven Tötung von Zellbeständen.
US4939666A (en) * 1987-09-02 1990-07-03 Genex Corporation Incremental macromolecule construction methods
AU4429989A (en) * 1988-10-19 1990-05-14 Dow Chemical Company, The A novel family of high affinity, modified antibodies for cancer treatment
WO1990006133A1 (fr) * 1988-11-25 1990-06-14 Centocor, Inc. Anticorps heterobifonctionnels se liant specifiquement a des plaquettes et a des agents thrombolytiques
DE3920358A1 (de) * 1989-06-22 1991-01-17 Behringwerke Ag Bispezifische und oligospezifische, mono- und oligovalente antikoerperkonstrukte, ihre herstellung und verwendung
GB9012995D0 (en) * 1990-06-11 1990-08-01 Celltech Ltd Multivalent antigen-binding proteins
ATE241011T1 (de) * 1991-02-27 2003-06-15 Micromet Ag Serin-reiche peptidlinker
CA2062582C (fr) * 1991-03-27 1996-03-26 Tse-Wen Chang Methodes et substances pour recruter des agents therapeutiques a des tissus solides
JPH07501451A (ja) * 1991-11-25 1995-02-16 エンゾン・インコーポレイテッド 多価抗原結合タンパク質
ES2102007T3 (es) * 1992-01-23 1997-07-16 Merck Patent Gmbh Proteinas de fusion de fragmentos de anticuerpo monomeras y dimeras.
CA2129663C (fr) * 1992-02-06 2005-07-05 James S. Huston Proteine fixatrice biosynthetique pour marqueur du cancer
US6329507B1 (en) * 1992-08-21 2001-12-11 The Dow Chemical Company Dimer and multimer forms of single chain polypeptides
DE69232604T2 (de) * 1992-11-04 2002-11-07 City Of Hope Duarte Antikörperkonstrukte
DK0672142T3 (da) * 1992-12-04 2001-06-18 Medical Res Council Multivalente og multispecifikke bindingsproteiner samt fremstilling og anvendelse af disse

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
INFECTION AND IMMUNITY, Volume 58, No. 5, issued May 1990, CLEMENTS, "Construction of a Non-Toxic Fusion Peptide for Immunization Against Escherichia Coli Strains that Produce Heat-Labile and Heat-Stable Enterotoxins", pages 1159-1166. *
JOURNAL OF THE NATIONAL CANCER INSTITUTE, Volume 82, No. 14, issued 18 July 1990, COLCHER et al., "In Vivo Tumor Targeting of a Recombinant Single Chain Antigen Binding Protein", pages 1191-1197. *

Cited By (56)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6824779B1 (en) 1993-07-26 2004-11-30 Dana-Farber Cancer Institute, Inc. Methods for inhibiting the interaction of B7-2 with its natural ligand
US7459544B2 (en) 1993-07-26 2008-12-02 Genetics Institute, Llc Nucleic acids encoding B7-2 fusion proteins
US6605279B2 (en) 1993-07-26 2003-08-12 Genetics Institute, Inc. Therapeutic compositions for inhibiting the interactions of B7-1 and B7-2 with their natural ligands
EP0839831B1 (fr) * 1994-02-18 2005-12-28 Washington University Gonadotropin monochaine
EP0839831A2 (fr) * 1994-02-18 1998-05-06 Sensi-Test Gonadotropin monochaine
US6103521A (en) * 1995-02-06 2000-08-15 Cell Genesys, Inc. Multispecific chimeric receptors
WO1996040915A3 (fr) * 1995-06-07 1997-01-23 Dana Farber Cancer Inst Inc Nouveaux ligands de ctla4/cd28 et leurs utilisations
EP1806402A3 (fr) * 1995-06-07 2007-09-19 Dana-Farber Cancer Institute, Inc. Ligands de CTLA4/CD28 et leurs utilisations
EP1806402A2 (fr) * 1995-06-07 2007-07-11 Dana-Farber Cancer Institute, Inc. Ligands de CTLA4/CD28 et leurs utilisations
WO1996040915A2 (fr) * 1995-06-07 1996-12-19 Dana-Farber Cancer Institute Nouveaux ligands de ctla4/cd28 et leurs utilisations
GB2333527A (en) * 1996-10-29 1999-07-28 Oxford Biomedica Ltd Therapeutic gene
GB2333527B (en) * 1996-10-29 2001-05-02 Oxford Biomedica Ltd Therapeutic gene
US6924123B2 (en) 1996-10-29 2005-08-02 Oxford Biomedica (Uk) Limited Lentiviral LTR-deleted vector
US6541219B1 (en) 1996-10-29 2003-04-01 Alan John Kingsman Therapeutic Gene
US7056699B2 (en) 1996-10-29 2006-06-06 Oxford Biomedia (Uk) Limited Lentiviral LTR-deleted vector
WO1998018934A1 (fr) * 1996-10-29 1998-05-07 Oxford Biomedica (Uk) Limited Gene therapeutique
US6743896B2 (en) 1997-04-30 2004-06-01 Enzon, Inc. Single-chain antigen-binding proteins capable of glycosylation, production and uses thereof
EP0979102A4 (fr) * 1997-04-30 2005-11-23 Enzon Inc Polypeptides a chaine unique modifies par oxyde de polyalkylene
US6743908B2 (en) 1997-04-30 2004-06-01 Enzon, Inc. Single-chain antigen-binding proteins capable of glycosylation, production and uses thereof
US6824782B2 (en) 1997-04-30 2004-11-30 Enzon, Inc. Polyalkylene oxide-modified single chain polypeptides
US6872393B2 (en) 1997-04-30 2005-03-29 Enzon, Inc. Polyalkylene oxide-modified single chain polypeptides
EP0979102A1 (fr) * 1997-04-30 2000-02-16 Enzon, Inc. Polypeptides a chaine unique modifies par oxyde de polyalkylene
EP0981548A4 (fr) * 1997-04-30 2005-11-23 Enzon Inc Proteines a chaine unique fixant les antigenes capables de glycosylation, production et utilisations de ces dernieres
EP0981548A1 (fr) * 1997-04-30 2000-03-01 Enzon, Inc. Proteines a chaine unique fixant les antigenes capables de glycosylation, production et utilisations de ces dernieres
US6323322B1 (en) 1997-04-30 2001-11-27 Enzon, Inc. Single-chain antigen-binding proteins capable of glycosylation, production and uses thereof
US7632504B2 (en) 1997-04-30 2009-12-15 Enzon, Inc. Polyalkylene oxide-modified single chain polypeptides
US7150872B2 (en) 1997-04-30 2006-12-19 Enzon, Inc. Polyalkylene oxide-modified single chain polypeptides
US6132722A (en) * 1997-05-07 2000-10-17 Bristol-Myers Squibb Company Recombinant antibody-enzyme fusion proteins
US6764853B2 (en) 1998-10-20 2004-07-20 Enzon Pharmaceuticals, Inc. Method for targeted delivery of nucleic acids
US6333396B1 (en) 1998-10-20 2001-12-25 Enzon, Inc. Method for targeted delivery of nucleic acids
WO2001023543A1 (fr) * 1999-09-24 2001-04-05 Large Scale Biology Corporation Creation de regions de liaison a sequence et longueur variables, pour molecules a domaine double ou multiple
US7297478B1 (en) 2000-09-22 2007-11-20 Large Scale Biology Corporation Creation of variable length and sequence linker regions for dual-domain or multi-domain molecules
EP1801121A1 (fr) * 2005-12-23 2007-06-27 CONARIS research institute AG Variantes solubles de la molécule gp130 utiles comme médicament
WO2007071449A1 (fr) * 2005-12-23 2007-06-28 Conaris Research Institute Ag Variantes de molécule gp310 solubles pouvant être employées en tant que médicament
US7851182B2 (en) 2005-12-23 2010-12-14 Conaris Research Institute Ag Soluble GP130 molecule variants useful as a medicament
US9573989B2 (en) 2006-06-30 2017-02-21 Conaris Research Institute Ag Sgp130/Fc dimers
US9034817B2 (en) 2006-06-30 2015-05-19 Conaris Research Institute Ag sgp130/Fc dimers for treatment of inflammatory disease
US8895012B2 (en) 2006-06-30 2014-11-25 Conaris Research Institute Ag sgp130/Fc dimers
US9624272B2 (en) 2013-03-14 2017-04-18 University Of Washington Through Its Center For Commercialization Polypeptides for treating and/or limiting influenza infection
WO2015165480A1 (fr) 2014-04-30 2015-11-05 Institute For Research In Biomedicine Compositions de vaccin de cytomégalovirus humain et leur procédé de production
WO2016033168A1 (fr) * 2014-08-26 2016-03-03 University Of Washington Through Its Center For Commercialization Polypeptides et leur utilisation pour le traitement de la grippe
US11198721B2 (en) 2014-12-01 2021-12-14 Ferring B.V. Methods for treatment of inflammatory disease or IL-6-mediated condition with gp130 protein
US10519218B2 (en) 2014-12-01 2019-12-31 Ferring B.V. Selective IL-6-trans-signalling inhibitor compositions
US11306136B2 (en) 2014-12-01 2022-04-19 Ferring B.V. Selective IL-6-trans-signalling inhibitor compositions
US11384350B2 (en) 2014-12-15 2022-07-12 The Regents Of The University Of California Cytotoxic molecules responsive to intracellular ligands for selective T cell mediated killing
WO2018013563A1 (fr) * 2016-07-12 2018-01-18 Kite Pharma, Inc. Molécules de liaison à l'antigène et procédés d'utilisation associés
US11608517B2 (en) 2016-07-12 2023-03-21 Kite Pharma, Inc. Antigen binding molecules and methods of use thereof
US10501775B2 (en) 2016-07-12 2019-12-10 Kite Pharma, Inc. Antigen binding molecules and methods of use thereof
CN109963590A (zh) * 2016-09-02 2019-07-02 加利福尼亚大学董事会 涉及白介素-6受体α结合单链可变片段的方法和组合物
WO2018042385A3 (fr) * 2016-09-02 2018-04-19 The Regents Of The University Of California Méthodes et compositions impliquant des fragments variables à chaîne unique (scfv) de liaison au récepteur alpha de l'interleukine-6
US11701384B2 (en) 2016-09-02 2023-07-18 The Regents Of The University Of California Methods and compositions involving interleukin-6 receptor alpha-binding single chain variable fragments
CN109963590B (zh) * 2016-09-02 2024-03-15 加利福尼亚大学董事会 涉及白介素-6受体α结合单链可变片段的方法和组合物
JP2020534030A (ja) * 2017-09-19 2020-11-26 パウル・シェラー・インスティトゥート トランスグルタミナーゼコンジュゲーション方法およびリンカー
WO2021108482A1 (fr) 2019-11-27 2021-06-03 The Board Of Trustees Of The University Of Illinois Pentapeptide et ses méthodes d'utilisation
WO2021236879A1 (fr) 2020-05-20 2021-11-25 The Board Of Trustees Of The University Of Illinois Méthode de traitement de maladies lysosomales à l'aide de peptides d'histatine
WO2023086787A1 (fr) * 2021-11-09 2023-05-19 Janssen Biotech, Inc. Dispositif et système de co-encapsulation microfluidique et procédés d'identification de ligands de récepteur de lymphocytes t

Also Published As

Publication number Publication date
AU5670194A (en) 1994-06-22
US5990275A (en) 1999-11-23
US5856456A (en) 1999-01-05

Similar Documents

Publication Publication Date Title
US5856456A (en) Linker for linked fusion polypeptides
Whitlow et al. An improved linker for single-chain Fv with reduced aggregation and enhanced proteolytic stability
JP3490437B2 (ja) 単量体および二量体抗体フラグメント融合タンパク質
Wels et al. Construction, bacterial expression and characterization of a bifunctional single–chain antibody–phosphatase fusion protein targeted to the human ERBB–2 receptor
EP0617706B1 (fr) Proteines multivalentes de fixation aux antigenes
Kipriyanov et al. Two amino acid mutations in an anti-human CD3 single chain Fv antibody fragment that affect the yield on bacterial secretion but not the affinity.
Whitlow et al. Single-chain Fv proteins and their fusion proteins
US6103889A (en) Nucleic acid molecules encoding single-chain antigen-binding proteins
ES2296395T3 (es) Construcciones de anticuerpos tetravalentes.
AU627183B2 (en) Method for producing recombinant dna proteins
Kipriyanov et al. High level production of soluble single chain antibodies in small-scale Escherichia coli cultures
Alfthan et al. Properties of a single-chain antibody containing different linker peptides
Eldin et al. High-level secretion of two antibody single chain Fv fragments by Pichia pastoris
Webber et al. Preparation and characterization of a disulfide-stabilized Fv fragment of the anti-Tac antibody: comparison with its single-chain analog
US20020076406A1 (en) Multivalent target binding protein
Li et al. Genetically engineered brain drug delivery vectors: cloning, expression and in vivo application of an anti-transferrin receptor single chain antibody–streptavidin fusion gene and protein
CA2135408A1 (fr) Analogues de proteines multivalentes chimeriques et modes d'utilisation
CA2793019A1 (fr) Anticorps humanises
Kurucz et al. A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor.
JP2001501801A (ja) モノクローナル抗体ｂｒ１１０およびその使用
CN101155832A (zh) 抗体的改良方法
De Jonge et al. Production and characterization of bispecific single-chain antibody fragments
George et al. Redirection of T cell-mediated cytotoxicity by a recombinant single-chain Fv molecule.
CN106478807B (zh) 识别mage-a3的t细胞受体
Bera et al. A bivalent disulfide-stabilized Fv with improved antigen binding to erbB2

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BR CA CZ FI HU JP KR NO NZ PL RO RU SK UA

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA